Cloning, expression and antimicrobial activity of crustinPm1, a major isoform of crustin, from the black tiger shrimp Penaeus monodon
Cloning, expression and antimicrobial activity of crustinPm1, a major isoform of crustin, from the black tiger shrimp Penaeus monodon
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DOI：
10.1016/j.dci.2007.04.004
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发表时间：
2008-01-01
影响因子：
2.9
通讯作者：
Tassanakajon, Anchalee
中科院分区：
文献类型：
--
作者：
Supungul, Premruethai;Tang, Sureerat;Tassanakajon, Anchalee
Crustin抗菌同系物，含有乳清酸性蛋白（WAP）结构域，已被确定从黑虎虾，斑节对虾血细胞库。这些cDNA的序列分析表明存在几种异构体的crustin在P monodon。CrustinPm 1是CrustinPm 1中含量最高的一种，其开放阅读框长435 bp，编码145个氨基酸的前体蛋白，其中包括17个氨基酸的信号肽和128个氨基酸的成熟肽。该肽在氨基端含有富含Gly-Pro的区域，在羧基端含有单个乳清酸性蛋白（WAP）结构域。为了表征该肽的性质和生物活性，在大肠杆菌中过量表达crustinPm 1。重组crustinPm 1的分子量为14.7kDa，预测pI为8.3。抑菌试验表明，重组crustinPm 1对革兰氏阳性菌具有较强的抑菌活性，对金黄色葡萄球菌和海豚链球菌具有较强的抑制作用。此外，抑菌机理研究表明，crustinPm 1的抑菌作用是杀菌作用的结果。原位杂交与crustinPm 1反义探针显示强杂交信号，在一定的血细胞群体的未受到挑战的虾，表明crustinPm 1转录差异表达在不同的血细胞亚群。(C)2007爱思唯尔有限公司保留所有权利。
Crustin antibacterial homologues, containing a whey acidic protein (WAP) domain, have been identified from the haemocyte library of the black tiger shrimp, Penaeus monodon. Sequence analysis of these cDNAs indicates the presence of several isoforms of crustin in P monodon. CrustinPm1, the most abundant isoform, contains an open reading frame of 435 bp encoding a precursor of 145 amino acids that comprises 17 amino acid signal peptides and 128 amino acid mature peptides. The peptides contain a Gly-Pro rich region at the amino-terminus and a single whey acidic protein (WAP) domain at the carboxyl-terminus. In order to characterize the properties and biological activities of this peptide, crustinPm1 was overexpressed in Escherichia coli. The recombinant crustinPm1 has a molecular mass of 14.7kDa with a predicted pl of 8.3. Antimicrobial assays demonstrated that recombinant crustinPm1 exhibited antimicrobial activity against only Gram-positive bacteria with strong inhibition against Staphylococcus aureus and Streptococcus iniae. In addition, the study of inhibition mechanism revealed that the antimicrobial activity of crustinPm1 was a result of bactericidal effect. In situ hybridization with crustinPm1 antisense probes showed strong hybridization signals in a certain haemocyte population of unchallenged shrimp, indicating that crustinPm1 transcript is differentially expressed in different subsets of haemocyte cells. (C) 2007 Elsevier Ltd. All rights reserved.